DETERMINATION OF COBALAMINS IN BIOLOGICAL-MATERIAL .2. THE COBALAMINS IN HUMAN-PLASMA AND ERYTHROCYTES AFTER DESALTING ON NON-POLAR ADSORBENT MATERIAL, AND SEPARATION BY ONE-DIMENSIONAL THIN-LAYER CHROMATOGRAPHY
DETERMINATION OF COBALAMINS IN BIOLOGICAL-MATERIAL .2. THE COBALAMINS IN HUMAN-PLASMA AND ERYTHROCYTES AFTER DESALTING ON NON-POLAR ADSORBENT MATERIAL, AND SEPARATION BY ONE-DIMENSIONAL THIN-LAYER CHROMATOGRAPHY
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DOI：
10.1016/0003-2697(83)90553-5
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发表时间：
1983-01-01
影响因子：
2.9
通讯作者：
HIPPE, E
中科院分区：
文献类型：
--
作者：
GIMSING, P;NEXO, E;HIPPE, E
描述了一种测定生物材料中钴胺素的改进方法。与醋酸镉一起预孵育后，用乙醇提取钴胺素，醋酸镉可抑制羟钴胺素对蛋白质的非特异性吸附。提取物在 Amberlite XAD-2 柱上脱盐，与之前的苯酚程序相比，分析能力至少增加了一倍。通过一维薄层色谱法分离钴胺素。在光敏钴胺素和羟钴胺素转化为磺钴胺素后，用大肠杆菌菌株 113-3 进行生物自显影。这样做是为了确保对等量的钴胺素的生长反应具有可比性。提供了健康献血者血浆和红细胞中甲基-、5''-脱氧腺苷-、氰基-、羟基/水-和磺基钴胺素的参考区间。讨论了当前结果和以前结果之间的差异。
An improved method for determination of cobalamins in biological materials is described. The cobalamins are extracted with ethanol after preincubation with cadmium acetate, which inhibits nonspecific adsorption of hydroxocobalamin to proteins. The extracts are desalted on Amberlite XAD-2 columns, which at least doubles the capacity of the analysis, as compared to the previous phenol procedure. The cobalamins are separated by 1-dimensional TLC. Bioautography with Escherichia coli strain 113-3 is performed after the light-sensitive cobalamins and hydroxocobalamin were converted to sulfitocobalamin. This is done in order to ensure comparable growth responses to equal amounts of the cobalamins. Reference intervals of plasma and erythrocytes of healthy blood donors for methyl-, 5''-deoxyadenosyl-, cyano-, hydroxo/aquo-, and sulfitocobalamin are presented. The differences between the present and previous results are discussed.